Ma Xuan, Wu Ming-xing, Zhang Yan-li, Cui Dong-mei, Li Ming-tao, Wu Kai-li
Zhongshan Ophthalmic Center, State Key Laboratory of Ophthalmology, Sun Yat-sen University, Guangzhou, Guangdong 510060, China.
Chin Med J (Engl). 2009 May 5;122(9):1072-80.
Equatorial lens epithelial cells proliferate and differentiate into fiber cells throughout life, while central lens epithelial cells proliferate little and do not form fiber cells. This study aimed to investigate the differences in gene expression between the central and the peripheral epithelial cells of the bovine lens.
Lens epithelia were dissected into central (<or= 11.5 mm diameter, cLEC) and peripheral regions (pLEC). The differences in gene expression and protein accumulation between these two regions were assayed by microarray analysis and two-dimensional electrophoresis (2-DE) combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Differently expressed proteins were validated by immunoanalyses.
By microarray analysis, 67 transcripts were at least two-fold lower and 269 at least two-fold higher in pLEC compared with that in cLEC. Thirty-four protein spots, including 20 in cLEC and 14 in pLEC, were identified by two dimensional electrophoresis and mass spectrometry. Of these 34 protein products, 28 were represented by probe sets on the microarray. Nine transcripts changed in the same direction and four transcripts in the opposite direction to their protein products. Immunoanalyses revealed that three (mitogen-activated protein kinase 1 (MAPK1), nidogen (NID), small nuclear ribonucleoprotein N (SNRPN)) out of four transcripts with opposite change between 2-DE and microarray assay showed the same changes as the results of 2-DE gel analyses. The genes differently expressed between cLEC and pLEC mainly include those related to the MAPK, transforming growth factor beta (TGFbeta) signaling and glycolysis pathways.
The results suggested that there were distinctly different genome activities, including a specific group of pathways, between central and peripheral lens epithelial cells.
赤道部晶状体上皮细胞在整个生命过程中增殖并分化为纤维细胞,而中央晶状体上皮细胞增殖很少且不形成纤维细胞。本研究旨在探讨牛晶状体中央和周边上皮细胞之间基因表达的差异。
将晶状体上皮分为中央区域(直径≤11.5mm,cLEC)和周边区域(pLEC)。通过微阵列分析以及二维电泳(2-DE)结合基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF-MS)检测这两个区域之间基因表达和蛋白质积累的差异。通过免疫分析验证差异表达的蛋白质。
通过微阵列分析,与cLEC相比,pLEC中67个转录本至少低两倍,269个至少高两倍。通过二维电泳和质谱鉴定出34个蛋白质斑点,其中cLEC中有20个,pLEC中有14个。在这34种蛋白质产物中,有28种由微阵列上的探针集代表。9个转录本与其蛋白质产物的变化方向相同,4个转录本与其蛋白质产物的变化方向相反。免疫分析显示,在2-DE和微阵列分析之间变化相反的4个转录本中,有3个(丝裂原活化蛋白激酶1(MAPK1)、巢蛋白(NID)、小核核糖核蛋白N(SNRPN))显示出与2-DE凝胶分析结果相同的变化。cLEC和pLEC之间差异表达的基因主要包括与MAPK、转化生长因子β(TGFβ)信号传导和糖酵解途径相关的基因。
结果表明,中央和周边晶状体上皮细胞之间存在明显不同的基因组活性,包括一组特定的途径。